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Antibodies, pharmaceutical compositions and methods

a technology of compositions and antibodies, applied in the field of antibodies, pharmaceutical compositions and methods, can solve the problems of breast cancer aggressiveness and poor prognosis

Active Publication Date: 2017-10-05
OBI PHARMA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present patent is about a newly discovered antibody that targets a protein called SSEA-4, which is found in high amounts in many types of cancer but is not present in normal cells. The antibody can be used as a diagnostic tool for detecting cancer and can also be used to treat the disease by targeting SSEA-4. The patent describes the structure of the antibody and its use in various applications.

Problems solved by technology

For example, Globo H has been shown to overexpress on a variety of epithelial cancers and is associated with tumor aggressiveness and poor prognosis in breast cancer and small cell lung carcinoma.

Method used

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  • Antibodies, pharmaceutical compositions and methods
  • Antibodies, pharmaceutical compositions and methods
  • Antibodies, pharmaceutical compositions and methods

Examples

Experimental program
Comparison scheme
Effect test

example 1

Fusion and Screening

[0376]A classical hybridoma fusion was performed. Mice received their first immunization with SSEA-4-DT-CRM197 (diphtheria toxin cross reacting material 197) conjugated with OBI-821 saponin adjuvant. Each mouse was administrated with designated treatment via subcutaneous (s.c.) injection at both left and right abdominal sites (100 μL / site). The mice were dosed four times (Day 0, Day 7, Day 14, and Day 23). Blood sample (approximately 0.1 mL whole blood / time point / animal) was obtained through facial vein without anticoagulant before Day 0 (pre-dose), Day 10, Day 17, Day 26 and on the sacrifice day (Day 35, volume of whole blood was collected through facial vein and heart punch). Coagulated blood samples were centrifuged (1500×g, 15 min, 4° C.), and serum was separated and transferred into eppendorf tubes. All serum samples were tested for titers of anti-SSEA-4 antibody. Five mice were found to produce high levels of anti-SSEA-4 IgG and IgM and were used for hybrid...

example 2

nt of the Anti-Tumor Activity of the Exemplary Antibody in Nude Mice

[0377]In a xenograft tumor model of human pancreatic adenocarcinoma, HPAC cells were subcutaneously (s.c.) implanted into BALB / c male nude mice and test articles were administered at 0.4 mg / kg by intraperitoneal (i.p.) injection twice weekly for 5 weeks starting when the average tumor size reached 50-120 mm3. The tumor size was monitored and recorded twice weekly for 36 days. Mortality, body weight, and signs of overt animal toxicity were also recorded twice weekly for 36 days. Tumor growth was calculated as T / C (treatment / control)×100%.

Test Substances and Dosing Pattern

[0378]Test substances were prepared at 1.45, 1, or 0.5 mg / mL in liquid form. Test substances were freshly prepared each day before dosing by diluting stocks solutions with phosphate buffer saline (PBS, pH7.4) to obtain the designated dosing concentration (0.04 mg / mL). Test substances were administered intraperitoneally (i.p.) at a dosing volume at 10...

example 3

tion of Cell Binding Ability of the Exemplary Antibody with MCF7 by FACS

Cell Culture

[0387]MCF-7 cell were cultured in Minimum essential medium (Invitrogen, Cat #10370021) with 2 mM L-glutamine (Invitrogen, Cat #25030081) and 1 mM sodium pyruvate (Invitrogen, Cat #11360070) and supplemented with 0.01 mg / mL insulin (Sigma, Cat #SI-I9278-5 mL), 10% fetal bovine serum (Invitrogen, Cat #16000044).

Cell Staining

[0388]Test cells were suspended by discarding media from the test sample flask containing monolayer cells. Cells were rinsed with 5 mL PBS twice. 1 mL of 0.05% trypsin was added to flask, swirled to cover all surface area, and put in a 37° C. CO2 incubator for 5-10 minutes. 5 mL of complete growth medium was added. Cells were aspirated by pipetting gently. Cell suspension was transferred to a 15 mL conical tube. The tube was centrifuged for 5 minutes at 200 g. Following centrifugation, the supernatant was removed, and the cell pellet was agitated.

Cell Counting

[0389]1-2 mL FACS buffe...

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Abstract

Pharmaceutical composition comprising antibodies or antigen binding fragments thereof that bind to stage-specific embryonic antigen 4 (SSEA-4) are disclosed herein, as well as methods of use thereof. Methods of use include, without limitation, cancer therapies and diagnostics. The antibodies of the disclosure can bind to certain cancer cell surfaces. Exemplary targets of the antibodies disclosed herein can include carcinomas, such as breast cancer, lung cancer, esophageal cancer, rectal cancer, biliary cancer, liver cancer, buccal cancer, gastric cancer, colon cancer, nasopharyngeal cancer, kidney cancer, prostate cancer, ovarian cancer, cervical cancer, endometrial cancer, pancreatic cancer, testicular cancer, bladder cancer, head and neck cancer, oral cancer, neuroendocrine cancer, adrenal cancer, thyroid cancer, bone cancer, skin cancer, basal cell carcinoma, squamous cell carcinoma, melanoma, and / or brain tumor.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims the priority of U.S. Provisional Patent Application No. 62 / 314,841, filed Mar. 29, 2016. The entirety of the aforementioned application is incorporated herein by reference.FIELD[0002]The present disclosure relates to antibodies and binding fragments thereof to carbohydrate antigens, as well as nucleic acids encoding such as antibodies, complementary nucleic acids, polypeptides, vectors, host cells, and methods of making and using thereof, including pharmaceutical compositions comprising said antibody and / or binding fragments. Furthermore, methods are provided for administering antibodies to a subject in an amount effective to inhibit cancer cells. Specifically, antibodies that bind to stage-specific embryonic antigen 4 (SSEA-4) are disclosed herein, as well as related compositions and methods of use. Methods of use include, without limitation, cancer therapies and diagnostics.BACKGROUND OF THE INVENTION[0003]Numero...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K16/18A61K49/00G01N33/574G01N33/92C12N5/16A61K39/395
CPCC07K16/18C07K2319/02A61K39/39558G01N33/57484G01N33/574A61K47/48569G01N33/92A61K49/00C07K2317/56C07K2317/565C07K2317/14C07K2317/24A61K2039/507G01N2400/00G01N2405/10C07K2317/31C07K2317/33C07K2319/03C12N5/16C07K16/44A61K2039/505A61P35/00G01N33/57438A61K49/0058C07K16/28C07K16/3076C12N5/163C07K2319/00
Inventor YU, CHENG-DER TONYLAI, JIANN-SHIUNCHEN, I-JULIN, CHIU-CHUN
Owner OBI PHARMA INC
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